title: Quantitative Segmentation of Fluorescence Microscopy Images of Heterogeneous Tissue: Application to the Detection of Residual Disease in Tumor Margins.
abstract: To develop a robust tool for quantitative in situ pathology that allows visualization of heterogeneous tissue morphology and segmentation and quantification of image features.
  
  
  
  TISSUE EXCISED FROM A GENETICALLY ENGINEERED MOUSE MODEL OF SARCOMA WAS IMAGED USING A SUBCELLULAR RESOLUTION MICROENDOSCOPE AFTER TOPICAL APPLICATION OF A FLUORESCENT ANATOMICAL CONTRAST AGENT: acriflavine. An algorithm based on sparse component analysis (SCA) and the circle transform (CT) was developed for image segmentation and quantification of distinct tissue types. The accuracy of our approach was quantified through simulations of tumor and muscle images. Specifically, tumor, muscle, and tumor+muscle tissue images were simulated because these tissue types were most commonly observed in sarcoma margins. Simulations were based on tissue characteristics observed in pathology slides. The potential clinical utility of our approach was evaluated by imaging excised margins and the tumor bed in a cohort of mice after surgical resection of sarcoma.
  
  
  
  Simulation experiments revealed that SCA+CT achieved the lowest errors for larger nuclear sizes and for higher contrast ratios (nuclei intensity/background intensity). For imaging of tumor margins, SCA+CT effectively isolated nuclei from tumor, muscle, adipose, and tumor+muscle tissue types. Differences in density were correctly identified with SCA+CT in a cohort of ex vivo and in vivo images, thus illustrating the diagnostic potential of our approach.
  
  
  
  The combination of a subcellular-resolution microendoscope, acriflavine staining, and SCA+CT can be used to accurately isolate nuclei and quantify their density in anatomical images of heterogeneous tissue.
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, North Carolina, United States of America.
authors.forename: Jenna L
authors.initials: JL
authors.lastname: Mueller
authors.forename: Zachary T
authors.initials: ZT
authors.lastname: Harmany
authors.forename: Jeffrey K
authors.initials: JK
authors.lastname: Mito
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Kennedy
authors.forename: Yongbaek
authors.initials: Y
authors.lastname: Kim
authors.forename: Leslie
authors.initials: L
authors.lastname: Dodd
authors.forename: Joseph
authors.initials: J
authors.lastname: Geradts
authors.forename: David G
authors.initials: DG
authors.lastname: Kirsch
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Willett
authors.forename: J Quincy
authors.initials: JQ
authors.lastname: Brown
authors.forename: Nimmi
authors.initials: N
authors.lastname: Ramanujam
mesh_terms: D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D018365:Neoplasm, Residual / Q000175:diagnosis* / Q000473:pathology